scholarly journals Molecular characterization of Aeromonas hydrophila isolates from diseased fishes in district Kasur, Punjab, Pakistan

2024 ◽  
Vol 84 ◽  
Author(s):  
L. Tooba ◽  
A. Shahzad ◽  
M. Zahid ◽  
R. Muhammad ◽  
I. Anam ◽  
...  
Keyword(s):  
Virulence Genes ◽  
Fish Farms ◽  
Biochemical Tests ◽  
Isolation Medium ◽  
Fish Samples ◽  
Causative Agents ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Epizootic Ulcerative Syndrome

Abstract Pakistan is an agricultural country and fisheries play a very important role in the economic development of the country. Different diseases are prevalent in Pakistani fish but information related to the causative agents is not well-known. Keeping in view the significance of bacterial pathogens as the causative agents of multiple fish diseases, the present study was conducted for identification, characterization and analysis of virulence genes of Aeromonas spp. isolated from diseased fishes. A total of fifty fish samples having multiple clinical indications were collected from different fish farms of district Kasur, Punjab Pakistan. For isolation of Aeromonas spp. samples were enriched and inoculated on Aeromonas isolation medium. Isolates were identified and characterized by different biochemical tests, Analytical Profile Index (API) 20E kit and Polymerase Chain Reaction (PCR) assays. All isolates were screened for three putative virulence genes including aerolysin (aer), haemolysin (hyl) and heat labile cytotonic enterotoxin (alt). Seven isolates of Aeromonas (A.) hydrophila were retrieved and identified based on API 20E. These isolates were further confirmed as A. hydrophila on the basis of PCR assays. Three isolates were detected positive for the presence of virulence genes (alt and hyl). Whereas aerolysin (aer) gene was not present in any of A. hydrophila isolates. The present study confirmed A. hydrophila as the causative agent of epizootic ulcerative syndrome and motile Aeromonas septicemia in fish farms of district Kasur, Punjab Pakistan. Moreover, detection of two virulence genes (alt and hyl) in A. hydrophila isolates is a threat for fish consumers of study area.

scholarly journals Molecular characterization of virulence factors in Aeromonas hydrophila obtained from fish

2012 ◽  
Vol 32 (8) ◽  
pp. 701-706 ◽  
Author(s):  
Samira T.L. Oliveira ◽  
Gisele Veneroni-Gouveia ◽  
Mateus M. Costa
Keyword(s):  
Virulence Factors ◽  
Aeromonas Hydrophila ◽  
Nile Tilapia ◽  
Virulence Genes ◽  
Saline Solution ◽  
Specific Primers ◽  
In Vivo Tests ◽  
Polymerase Chain

Multiple factors can be involved in the virulence processes of Aeromonas hydrophila. The objective of the present paper was to verify the presence of aerolysin, hidrolipase, elastase and lipase virulence genes through the polymerase chain reaction (PCR) in A. hydrophila isolates obtained from fish of the São Francisco River Valley, and to evaluate virulence according to the presence of these genes in Nile tilapia fingerlings. One hundred and fourteen isolates from the bacteria were used. DNA was heat extracted and PCR undertaken using specific primers described in the literature. For in vivo tests Nile tilapia fingerlings were used. From the PCR tests, negative isolates for all genes tested were selected, positive isolates for two genes (aerolysin and elastase) and positive for the four genes tested. These were inoculated at a concentration of 10(8) UFC/ml into the tilapias, considered as treatments; another group of animals was used as control (with inoculation of saline solution). In all, 12 distinct standards regarding the presence of virulence factors in isolates from A. hydrophila, were observed. Of the 114 isolates analyzed, 100 (87.72%) presented at least one of the virulence factors under study. The virulence factors were widely distributed among the A. hydrophila isolates. Aerolysin was the most frequent virulence factor present in the isolates analyzed. A. hydrophila led to the mortality of the Nile tilapia fingerlings, regardless of the absence or quantity of virulence genes tested.

scholarly journals Detection and Molecular Characterization of Vibrio Parahaemolyticus in Shrimp Samples

2018 ◽  
Vol 12 (1) ◽  
pp. 46-50 ◽  
Author(s):  
Daryoush Asgarpoor ◽  
Fakhri Haghi ◽  
Habib Zeighami
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Vibrio Parahaemolyticus ◽  
Foodborne Diseases ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Global Issue ◽  
Retail Outlets ◽  
Polymerase Chain

Background:Food safety has emerged as an important global issue with international trade and public health implications. Bacterial pathogens asVibrio parahaemolyticusrecognized as an important cause of foodborne diseases related to the consumption of raw, undercooked or mishandled seafood worldwide.Methods:A total of 70 individual wild shrimp samples were collected from shrimp retail outlets in Zanjan, Iran and investigated for the presence of potentially pathogenic strains ofV. parahaemolyticus.The shrimp samples were immediately homogenized and cultured on TCBS agarand subjected to confirmatory biochemical tests. Polymerase Chain Reaction (PCR) was performed for detection of total and pathogenicV. parahaemolyticusby amplification ofvp–toxR,tdhandtrhgenes.Results:The conventional method indicated that 16 (22.8%) of samples were positive forV. parahaemolyticus. However, PCR verified that only 12 (17.1%) shrimp samples were positive forV. parahaemolyticus.Of the 70 shrimp samples in our study, only 2 (2.8%)tdhand 1 (1.4%)trhpositive strains were identified.Conclusion:Detection oftdhand/ ortrhpositiveV. parahaemolyticusin shrimp marketed in Zanjan, Iran shows a probable risk for public health. Therefore, the reliable molecular methods for monitoring of potentially pathogenicV. parahaemolyticusare strongly recommended for the routine seafood examination.

scholarly journals Genotypical characterization of thermotolerant coliforms isolated from food produced by a Solidarity Economic Venture of Bahia (Brazil)

2020 ◽  
Author(s):  
J. N. Silva ◽  
M. D. Baliza ◽  
F. Freitas ◽  
E. S Cruz ◽  
V. M. A. Camilo ◽  
...  
Keyword(s):  
Virulence Genes ◽  
Food Contamination ◽  
Microbiological Analysis ◽  
Chain Reaction ◽  
E Coli ◽  
Family Farmers ◽  
Two Samples ◽  
Thermotolerant Coliforms ◽  
Polymerase Chain

Abstract Many Solidarity Economic Venture (SEV) are family farmers who seek to add value to production through artisanal processing, which can lead to food contamination. Thus, this study aimed to genotypically characterize thermotolerant coliforms (TtC) strains from food produced by local agribusinesses of SEV during January to April 2019. Samples from thirteen production units (PU) from the SEV were submitted to a microbiological analysis of thermotolerant coliforms (AFNOR 3M1/2 – 09/89), using a fast count method in Petrifilm™ dishes. The Polymerase Chain Reaction (PCR) technique was used to verify the following virulence genes (VGs) associated with Escherichia coli: stx, typical from enterohemorrhagic E. coli (EHEC); bfpA typical from entheropathogenic E. coli (EPEC) and elt and slt, typical from entherotoxigenic E. coli (ETEC). The results showed that two samples of queijadinha (typical Brazilian candy made with eggs and coconut) and one sample of cassava cake presented characteristic colonies TtC. This way, three strains were isolated in order to perform the PCR technique. However, the genes used in the reaction were not detected in the isolated strains. Therefore, it is suggested that the isolated strains are from E. coli pathotypes with different virulence genes than the ones analyzed belong other types of TtC, such as Enterobacter and Klebsiella. Although the virulence of genes has not been confirmed, the presence of TtC on food indicates hygiene flaws during production and, therefore, measurements to control and prevent contamination should be taken.

scholarly journals Genetic Differentiation of Clariid Populations using Microsatellite Markers in Kano State Rivers

2021 ◽  
pp. 35-45
Author(s):  
I. O. Suleiman ◽  
R.O. Okeke ◽  
J. M. Madu ◽  
A. U. Umar ◽  
O.M Akinsola ◽  
...  
Keyword(s):  
Genetic Variation ◽  
Genetic Differentiation ◽  
Dna Extraction ◽  
Genetic Characterization ◽  
Caudal Peduncle ◽  
Fta Cards ◽  
Fish Samples ◽  
Polymerase Chain ◽  
Outbred Populations

This study aimed to investigate the genetic characterization of strains of Clariid fish species in some river bodies in Kano State using microsatellite markers.One hundred and seventy seven Clariid fish samples (Clariasgariepinus and Heterobranchuslongifilis) were collected from six rivers (Thomas, Ghari, Tiga dam, Duddurun Gaya, Karaye and Bagwai) in Kano state. Blood sample was taken from each fish sample by severing the caudal peduncle and drained into FTA cards for DNA extraction, Polymerase Chain Reaction and electrophoresis to determine genetic variation between the Clariid fish populations.Genealex 6.4 software package was used to analyse the resolve bands from DNA extraction to determine their base pair and genetic variation. Results showed that the Fst values ranged from 0.00 to 0.66, Fit ranged from -0.04 to 0.12, Fis ranged from -0.35 to -0.26. It indicated a large number of gene flow (exchange) among the populations with a range of 0.46 to 0.87. There was an established magnitude of genetic divergence (91.86%) among the populations as shown by the result of the percentage polymorphism which depends on the number of alleles detected per locus and their frequencies. It can be concluded that since there was no inbreeding as shown in the study, none of the population exhibited genetic uniqueness. The populations had a high genetic differentiation between populations but moderate differentiation within populations. The populations were outbred populations; an indication that relatives avoided mating in the population.

scholarly journals Virulence-Associated Gene Profiles of Avian Pathogenic Escherichia coli Isolated From Broilers With Colibacillosis: A Pilot Study in Iran

2019 ◽  
Vol 7 (1) ◽  
pp. 4-8
Author(s):  
Payam Haghighi Khoshkhoo ◽  
Hadi Pourtaghi ◽  
Gita Akbariazad ◽  
Saeed Mokhayeri
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Economic Losses ◽  
Avian Pathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Gene Profiles ◽  
Pathogenic Escherichia Coli ◽  
Poultry Farms ◽  
Polymerase Chain

Background: Avian pathogenic Escherichia coli (APEC) causes economic losses in the chicken industry worldwide. Objective: In this study, virulence-associated gene profiles of APEC isolates were investigated by polymerase chain reaction (PCR). Materials and Methods: A total of 60 Escherichia coli isolates were collected from 60 colibacillosis cases from 30 broiler poultry farms in Alborz, Tehran, and Golestan provinces, Iran. After identification by biochemical tests, DNA was extracted by boiling method and 5 virulence-associated genes including: iutA, hlyF, iroN, ompT, and iss were detected by 2 multiplex PCR protocols. Results: Of the 60 APEC isolates, 26 (43.3%) isolates had at least three virulence genes from which 12 (20%) isolates were positive for all 5 virulence genes, whereas 34 (56.6%) carried no investigated virulence genes. Presence of iutA, hlyF, iroN, ompT, and iss genes in the APEC isolates were 17 (28.3%), 17 (28.3%), 24 (40%), 26 (43.3%), and 23 (38.3%), respectively. Conclusion: According to the results, four different virulence-associated gene profiles were seen in isolates, from which profile 1 with 12 (20%) isolates was predominant. These findings were in agreement with the previous reports.

scholarly journals Identification of a 16SrII-E Phytoplasma in Calendula arvensis, Solanum nigrum, and Chenopodium spp.

Plant Disease ◽  
2006 ◽  
Vol 90 (3) ◽  
pp. 325-330 ◽  
Author(s):  
G. Tolu ◽  
S. Botti ◽  
R. Garau ◽  
V. A. Prota ◽  
A. Sechi ◽  
...  
Keyword(s):  
Solanum Nigrum ◽  
Nested Polymerase Chain Reaction ◽  
Bois Noir ◽  
Gene Coding ◽  
Epidemiological Surveys ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Taxonomic Groups ◽  
Stolbur Phytoplasma

Epidemiological surveys were performed in Northern Sardinia (Italy) in a 10-year-old vineyard affected by “Bois noir” disease. Samples collected between May and October 2003 from chlorotic and stunted weeds belonging to 14 different taxonomic groups were indexed molecularly for detection of phytoplasmas. Nested polymerase chain reaction (PCR) assays using primers specific for the phytoplasma 16SrDNA gene showed three of six Calendula arvensis, one of two Solanum nigrum, and one of seven Chenopodium spp. assayed positive. Restriction fragment length polymorphism analyses and sequencing of amplified 16SrDNA fragments identified a putative phytoplasma in the ribosomal subgroup 16SrII-E. Further characterization of the rps3 gene, coding a ribosomal protein, confirmed the identification. However, the weeds and leafhop-per species collected in the vineyard tested negative by PCR assays for the Stolbur phytoplasma, the causal agent of “Bois noir”. This is the first report of a phytoplasma of the 16SrII-E subgroup infecting C. arvensis, S. nigrum, and Chenopodium spp.

scholarly journals Characterization of phytoplasmas related to 'Candidatus Phytoplasma asteris' subgroup rpI-L in Iran

2014 ◽  
Vol 54 (2) ◽  
pp. 199-203 ◽  
Author(s):  
Fereshteh Vali-Sichani ◽  
Masoud Bahar ◽  
Leila Zirak
Keyword(s):  
Fragment Length Polymorphism ◽  
Specific Primer ◽  
Sequence Analyses ◽  
Chain Reaction ◽  
Universal Primer ◽  
Disease Symptoms ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Niger Seed

Abstract In two of Iran's central provinces, several herbaceous plants showing phytoplasma disease symptoms were collected to detect 'Canididatus Phytoplasma asteris'-related phytoplasmas. Confirmation of an association of phytoplasmas with diseased plants was done using polymerase chain reaction (PCR) assays having the phytoplasma universal primer pairs P1/P7 followed by R16F2n/ R16R2 in nested PCR. Then, for detection of 'Ca. P. asteris', DNA samples were subjected to amplification of rp and tuf genes using specific primer pairs rp(I)F1A/rp(I)R1A and fTufAy/rTufAy, respectively. Restriction fragment length polymorphism or RFLP analyses of rp gene fragments using Tsp509I restriction enzyme as well as sequence analyses indicated that 'Ca. P. asteris'-related phytoplasmas associated with carrot, niger seed and scallion plants in these regions, belong to the rpI-L subgroup. This research is the first report of carrot, niger seed, and scallion infection with phytoplasmas belonging to the rpI-L subgroup.

scholarly journals Molecular Characterization of Carbapenemase-Producing Escherichia coli and Salmonella in children with diarrhea in rural Burkina Faso

2020 ◽  
Author(s):  
Rene DEMBELE ◽  
Issiaka Soulama ◽  
Wendpoulomdé A. D. Kaboré ◽  
Ali Konaté ◽  
Assèta Kagambèga ◽  
...  
Keyword(s):  
Burkina Faso ◽  
Treatment Options ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Resistance Rates ◽  
Encoding Genes

Abstract Background: In recent years, carbapenemase-producing Enterobacterales (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso. Methods: Salmonella isolates were serotyped according to the Kauffman White scheme. Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were carried out using oligonucleotides to detect the presence of genes of the blaKPC, blaVIM, blaIMP, blaTEM, blaSHV, blaOXA and blaCTX-M types in all E. coli and Salmonella strains.Results: The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E. coli were imipenem resistant with carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each in E. coli strains. However, no Salmonella carbapenemases blaKPC, blaVIM or blaIMP were detected.Conclusions: This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacterales in patients is crucial.

scholarly journals Antibiotic Susceptibility Pattern, Molecular Characterization of Virulence Genes among Pseudomonas Aeruginosa Isolated from Burn Patients

2021 ◽  
Vol 5 (1) ◽  
pp. 36-41
Author(s):  
Mustafa D. Younus ◽  
Omar F. Bahjat ◽  
Sirwan A. Rashid
Keyword(s):  
Virulence Genes ◽  
Antimicrobial Agents ◽  
Burn Patients ◽  
Chain Reaction ◽  
The West ◽  
Antibiotic Susceptibility Pattern ◽  
Amoxicillin Clavulanic Acid ◽  
Polymerase Chain ◽  
Biochemical Testing

Abstract In this research a total of 150 samples were obtained from burn and wound patients admitted to the West Erbil Emergence Hospital during period from September 2020 to January 2021. Through cultural, morphological features, biochemical testing and Vitek’s 2 compact systems, 40 isolates of P. aeruginosa have been identified. P. aeruginosa produced various pigments, including blue / green, and yellow / green. The iso1ates of P. aeruginosa were subjected to 14 different antibiotics. Impenim was the most effective antimicrobial agents against all P. aerugionsa isolates, and most of isolates showed high resistance degree to Ampicillin 100%, Chloramphenicol 100%, amoxicillin-clavulanic acid 100%, Cefotaxime 100% and Penicillin 100% while for Aztreonam 32.5%, Meropenem 42.5%, Tobramycin 45%, Gentamycin 45%, Amikacin 45%, Ciprofloxacillin 62.5%, ceftazidime 67.5, % Tetracycline 80%. All Psudomonas aeruginosa isolates were screened using Multiplex polymerase chain reaction (PCR) to check for the presence of (Pvda, LasB, Protease, exoA, exoT, exoU and plch) on its genomic DNA. The findings have shown that (Pvda was 55%, LasB 75%, Protease 65%, exoA 60%, exoT 75%, exoU 60% and, plch 55%) of isolates harbored these genes as a virulence genes.

scholarly journals Partial Molecular Characterization of Dahlia mosaic virus and Its Detection by PCR

Plant Disease ◽  
2003 ◽  
Vol 87 (8) ◽  
pp. 945-948 ◽  
Author(s):  
M. Nicolaisen
Keyword(s):  
Nucleotide Sequence ◽  
Mosaic Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Open Reading Frames ◽  
Dahlia Mosaic Virus ◽  
Polymerase Chain ◽  
Pcr Assays

Dahlia mosaic virus (DMV) is the causal agent of one of the most important diseases of Dahlia pinnata. The nucleotide sequence of a 1,195-bp fragment of its genome was amplified and characterized. Based on this sequence, polymerase chain reaction (PCR) assays were developed for detection of DMV. The nucleotide sequence confirmed the classification of DMV as a member of genus Caulimovirus since it was similar to a region covering partly open reading frames (ORFs) IV and V found in caulimoviruses. The two most closely related viruses on the basis of comparison of ORF V fragments were shown to be Figwort mosaic virus and Mirabilis mosaic virus with 66.6 and 68.1% identity, respectively. Two PCR assays were developed using identical primer pairs: a real-time PCR based on SYBR green chemistry and a conventional PCR. Both methods clearly discriminated DMV-infected and healthy dahlia. The real-time PCR assay detected DMV-infected material that was diluted 105-fold in healthy material.

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